Please use this identifier to cite or link to this item: http://hdl.handle.net/11455/53558
標題: 基因轉殖纖維/木質纖維分解微生物之應用
Application of the Gene-Transformed Xylanolytic/Cellulolytic Microbes
作者: 林瑞文
關鍵字: 生物技術;應用研究;Cellulolytic/Xylanolytic Microbes;纖維分解菌;纖維分解基因;基因轉殖纖維分解微生物;cellulase or Xylanase Gene;Lux Gene-transformed Cellulolytic Microbe
摘要: 
纖維之利用在產業界是多元的,造紙、飼料、肥料、食品加工皆可為目標產業項目。分離、篩選不同類型纖維分解菌供利用為一可行方法,惟大部分微生物皆具潛在病原性,纖維分解菌之利用亦可能受限於其潛在病原性或其生長、生理條件限制。選殖利用特定纖維分解基因 cellulase or xylanase genes 可以更具實際應用的可行性。選殖特定纖維分解基因加以改造、轉殖於特定微生物為 ”基因轉殖纖維分解微生物 Gene-Transformed Cellulolytic/Xylanolytic Microbes” 再加利用,更具安全性與可控制性。”基因轉殖纖維分解微生物” 可以直接利用,或亦可利用其生產纖維分解酵素 Cellulases or Xylanases 再利用。目前已篩選 X. campestris 之纖維分解基因 cellulase genes Xcc0027, Xcc0028, Xcc1752, Xcc3521, Xcc3381 等,轉殖於 E. coli與 X. campestris 證明此等基因皆具纖維分解功能。研究已建立有關基因轉殖模式。Lux-Gene-Transformed Cellulolytic/Xylanolytic Microbes 其 reporter genes 螢光基因亦可表現、並可加以追蹤分析,此轉殖模式可作纖維分解基因轉殖工具。延續計劃之研究構想:(1) 選殖 X. campestris xylanase genes 利用已建構之轉殖模式,構築Gene-Transformed Xylanolytic Microbes,以分析研究 Xylanase Genes Xcc0144, Xcc0855, Xcc4115, Xcc4118 之木質纖維分解酵素之功能。(2) 改造、強化基因表現系統以促進 Gene-Transformed Cellulolytic/Xylanolytic Microbes 之纖維/木纖維分解能力;已取得P. leiognathi R&R[HI] 與 Xcc phage ?烴7 R&R[?烴] 強化表現之調節區域可供利用。(3) 繼續篩選、分離纖維分解基因。(4) 建構適當的 Lux-shuttle vectors or broad-host range vectors 供作轉殖工具,建立轉殖方法,轉殖纖維分解基因至相關無病原性菌種,建構 ”基因轉殖纖維分解微生物”。(5) 分析、評估 ”基因轉殖纖維分解微生物” 之纖維分解能力。(6) 衡量纖維分解基因與固氮基因、溶磷基因、蛋白質分解基因共存之可能性,開發多功能之基因轉殖菌種 Labile-Programmed Lux-Gene-Transformed Cellulolytic/Xylanolytic Microbes;(7) 瞭解纖維分解菌之纖維分解功能機制,並偵測、評估外加纖維分解菌追蹤其對作物生長的效益;(8) 評估 ”基因轉殖纖維分解微生物” 作為組合複方生物製劑之可行性,亦探討病原性的預防。

Cellulases or cellulolytic microbes are potentially useful for application in agriculture, industry or food processing. Cellulolytic microbes are able to degrade cellulose to particular product. However, most of the native cellulolytic microbes are potential pathogenic, somehow it is dangerous in application. Thus, to clone various cellulase or xylanase genes, and transform these genes to the various non-pathogenic microbes would be useful in the future. The specific cellulases or xylanases from the gene-transformed cellulolytic microbes will be produced. Also it is possible to apply the gene-transformed cellulolytic microbes directly. Based on the concept and idea, the specific cellulase or xylanase genes will be cloned and selected, and to construct the Lux-gene-transformed cellulolytic microbes. Xanthomonas campestris cellulase genes Xcc0027, Xcc0028, Xcc1752, Xcc3521, Xcc3381 have been cloned, and transformed into E. coli and X. campestris. The result demonstrates that all the five Lux-gene-transformed cellulolytic E. coli indeed function to produce and secrete the cellulases to degrade the cellulose. The Lux-gene- transformed cellulolytic E. coli perform various cellulase functions. It will be able to combine various Lux-transformed cellulolytic microbes for the specific purpose application. Meanwhile, the luxA-luxB genes of luciferase are used as reporter genes for the cellulase or xylanase gene expression, also bioluminescence will be functioned as tracing marker. The Lux-cellulase genes or xylanase genes will be transformed into non-pathogenic, food-safe microbes, i.e. Bacillus, Xanthomonas etc. The Lux-gene- transformed cellulolytic microbes will be able to apply in food procession and the related procession. The project plan will continue to clone xylanase genes Xcc0144, Xcc0855, Xcc4115, Xcc4118, and to make the Lux-gene-transformed xylanolytic microbes. It is expected that the gene-transformed cellulolytic/ xylanolytic microbes will be able to form mixed combined complex bio-reagents for application.
URI: http://hdl.handle.net/11455/53558
其他識別: 96農科-1.2.1-科-a1(13)
Appears in Collections:生物化學研究所

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